Since the first commercial approval in the 1980s, dozens of monoclonal antibodies (mAbs) have been used for therapeutic use, including treatments for cancer, autoimmune diseases, and infectious disease. While mAbs’ specificity and affinity have made them popular targets for biopharmaceutical development, they are considerably more expensive and complicated to develop and manufacture than small molecules.
Patient safety is a priority when developing and manufacturing mAbs, and stringent regulatory guidelines must be followed to ensure the therapeutics are of sufficient purity. Purification of mAbs is routinely performed using protein A based affinity chromatography, which provides high capacity and consistent purification performance. When using protein A-based techniques, regulatory bodies outline requirements for quantification of the leached protein A ligand in the downstream material.
For the most accurate measurement of ligand leakage, it is critical that manufacturers use a resin-matched standard to calibrate their assay. A newly available, enzyme-linked immunosorbent assay from Cytiva is helping biopharmaceutical labs using MabSelect PrismA™ chromatography resin and Fibro™ PrismA chromatography adsorbers measure ligand leakage to meet regulatory requirements for purity data.
The PrismA ELISA kit was developed with a focus on usability, robustness, and environmental sustainability. The kit contains all the necessary reagents, including the PrismA ligand optimized for use in ELISA and polyclonal antibodies raised specifically against the PrismA ligand. The matched PrismA protein A ligand included in the kit removes the need to source the standard separately and create a custom protocol using a generic protein A assay. Its parts per billion sensitivity minimizes the risk of undetected residual PrismA ligand, and high IgG tolerance simplifies sample preparation.
Downstream samples generally have high concentrations of IgG, which can make measuring protein A a challenge. This is because accurate measurement requires disassociation of the protein A ligand and IgG, which is especially difficult when the concentration of the ligand is very low (< 1 ppm). When high, medium, and low concentrations of PrismA ligand were spiked into decreasing concentrations of IgG, the PrismA ELISA kit showed accurate recovery with as high as 20 mg/mL of IgG in the sample. The PrismA ELISA kit is also capable of detecting low amounts of ligand achieving single parts per billion (ppb) detection, up to and including the final stages of purification.
The PrismA ELISA kit also offers consistently low intra- and inter-plate variability to ensure reproducible data, and wide buffer compatibility removes the need for complicated and time-consuming buffer-exchange protocols that may compromise the integrity of samples. A broad dynamic range also helps to reduce the time and guesswork needed to dilute samples into the assay range.
The kit’s attention to detail even extends to packaging, which emphasizes recyclable materials and paper, and provides instructions via QR code.
In summary, Cytiva’s new PrismA ELISA kit accelerates a key step in mAb development and will help biopharmaceutical labs bring life-saving therapeutics to patients faster.